Three kinds of affinity membranes utilized for endotoxin removal, namely, chitosan affinity membrane (KFCC517), hydrophobic and cationic charged membrane (KFCG316) and metal chelate affinity membrane (KFCM402) were prepared by using cellulose membrane as support. The adsorbability and applications of the membranes in endotoxin removal were investigated. The results showed that all of these affinity membranes could be utilized for endotoxin removal in several kinds of solutions. The factors affecting endotoxin removal such as ionic strength, pH value and flow rate were investigated, and the results were discussed. Three cartridges of membranes KFCC517, KFCG316 and KFCM402 were utilized for endotoxin removal in human serum albumin solution. The cartridge of membrane KFCC517 also was utilized in medicine injections, such as hydrocortisone, puerarin, tetracaine hydrochloride and dextran 40 glucose. Under the optimal conditions, the removal efficiencies of endotoxin were over 80% and the recoveries of desirable substances were over 95%.